The document discusses the process of translation, where mRNA is used to synthesize proteins from amino acids. Translation occurs through three main stages - initiation, elongation, and termination - and involves various tools like amino acids, mRNA, tRNAs, ribosomes, and other factors. While similar between prokaryotes and eukaryotes, translation differs in some initiation and elongation factors used, and eukaryotic mRNA contains a 5' cap and 3' poly-A tail.

1. M S . N E H A
A s s t . P r o f . ( Z o o l o g y )
Protein Synthesis
(Translation)

2. What is translation?
 Formation of protein from mRNA is called translation, is
also known as polypeptide synthesis or protein synthesis. It
is unidirectional process.
 The raw materials for protein synthesis are amino acids,
mRNA, tRNAs and amino acyl tRNA synthetases.
 Ribosomes catalyze the joining of the amino acid
monomers directed by the mRNA sequence.
 Amino-acyl tRNA synthetases attach amino acids to the
appropriate tRNAs.
 The amino-acyl tRNA act as adaptors in the translation of
the nucleic acid sequence of the mRNA into the amino acid
sequence of the protein.

3. Translation occurs in three stages...
 Initiation- The components of the translational
apparatus come together with the mRNA molecule (A tRNA
carrying the first amino acid binds to the start codon).
 Elongation- Amino acids are brought to the mRNA as
amino-acyl tRNAs and are added one at a time to a growing
polypeptide chain.
 Termination- A stop codon in the mRNA is recognized by
a protein release factor and the translational apparatus
comes apart to release a completed polypeptide.

4. Tools of Translation…
 Amino acids : Twenty types of amino acids and amides constitute the
building blocks of proteins.
 mRNA : It carries the coded information for synthesis of one
(unicistronic) or more polypeptides (polycistronic). Its codons are
recognised by tRNAs
 tRNAs : They picks up specific amino acid from amino acid pool and
carrying over the mRNA strand.
 Amino Acyl tRNA Synthetases : The enzymes are specific for particular
amino acids and their tRNAs.
 Ribosomes : The A (aminoacyl) and P (peptidyl) sites as cavities on the
ribosome where charged tRNA (carrying an amino acid) molecules bind
during polypeptide synthesis. The recently postulated E (exit) site is the
site from which discharged tRNAs leave the ribosome. The mRNA-binding
site binds a sequence near the 5 prime end of the mRNA.
The binding sites are all located at or near the interface between the large
and small subunits

6. Charging of tRNA
Step 1) The amino acid and a
molecule of ATP enter the active
site of the enzyme.
The ATP loses pyrophosphate
and the resulting AMP bonds
covalently to the amino acid. The
pyrophosphate is hydrolyzed into
two phosphate groups.
Step 2) The tRNA covalently
bonds to the amino acid to
displace the AMP and the
aminoacyl tRNA is then released
from the enzyme

7. The Process of Translation
 A prokaryotic mRNA molecule encoding a single polypeptide has 5
prime non-coding leader sequence which contains a ribosome binding
site
 The rest of the mRNA contains a coding sequence that starts with an
AUG start codon and ends with a stop codon (UAA, UAG or UGA) and a
3 prime non-coding trailer sequence
 A eukaryotic mRNA molecule has, in addition to the above, a 5 prime
cap and a 3 prime poly(A) tail. One important difference is that
eukaryotic mRNAs lack a ribosome binding site (SD site)
 N-formylmethionine (fMet) is the modified amino acid with which
every polypeptide is initiated in prokaryotes but methionine is the
first amino acid in eukaryotes

8. Translation in prokaryotes
Initiation
Formation of the 70S translation
initiation complex occurs in
three steps:-
 Three initiation factors (IF) and
GTP bind to the small ribosomal
subunit.
 The initiator aminoacyl tRNA and
mRNA are attached. The mRNA-
binding site is composed of a
portion of the 16S rRNA of the
small ribosomal subunit. The 3’
end of the 16S rRNA bears a
pyrimidine-rich stretch that base
pairs with the Shine-Dalgarno
sequence of the mRNA)
 The large ribosomal subunit joins
the complex. The resulting 70S
initiation complex has fMet-
tRNAfMet residing in the ribosome's
P site

9. Chain elongation
• Elongation begins with the binding of the
second aminoacyl tRNA at the ribosomal
aminoacyl (A) site. The tRNA is escorted to
the A site by the elongation factor EF-Tu,
which also carries two bound GTPs. As the
tRNA binds, the GTPs are hydrolyzed and
EF-Tu is released. EF-Ts helps recycle the
EF-Tu.
• A peptide bond is formed between the
carboxyl group of the terminal amino acid
(or fMet in the first cycle) at the P site and
the amino group of the newly arrived
amino acid at the A site. This reaction is
catalyzed by the peptidyl transferase
activity of the 23S rRNA molecule in the
large ribosomal subunit.
• After EF-G-GTP binds to the ribosome and
GTP is hydrolyzed, the tRNA carrying the
elongated polypeptide translocates from
the A site to the P site. The discharged
tRNA moves from the P site to the E (exit)
site and leaves the ribosome. As the
peptidyl tRNA translocates, it takes the
mRNA along with it. Consequently, the
next mRNA codon is moved into the A site,
which is open for the next aminoacyl
tRNA. These events are repeated for each
additional amino acid.

10. Termination
Depends on release factors that recognize the three stop codons. When a stop codon (UAG,
UAA, or UGA) arrives at the A site, it is recognized and bound by a protein release factor. This
protein causes the polypeptide to be transferred to a molecule of water to cause its release
from the tRNA and the dissociation of the other components of the elongation complex.

11. Translation in Eukaryotes
Initiation
 In eukaryotes, translation
includes a different set of
initiation factors (eIFs), a
slightly different assembly
pathway
 eIF2 binds the initiator
tRNA met before the small
ribosomal subunit. This
complex can attach to the 5
prime cap structure of the
mRNA.
 As there is no Shine-Dalgarno
sequence, the ribosome begins
translation at a AUG that is
located within the Kozak
consensus sequence (often a
good match to CAAAAUG).

12. Chain elongation
 The process of elongation in eukaryotes is same as in
prokaryotes
 But elongation factors are different- eEF1 and eEF2
in eukaryotes.
Termination
 Termination factor eRF1 is required for termination
in eukaryotes
 Procedure is same as in prokaryotes